INTRODUCTION: Children with Attention Deficit/Hyperactivity Disorder (ADHD) participated in a randomized clinical trial comparing animal-assisted intervention (AAI) to psychosocial treatment as usual (TAU). This brief report describes effects of AAI on acute HPA axis reactivity and regulation. Saliva was collected before, during, and after psychosocial intervention sessions with and without therapy dogs and later assayed for cortisol (ug/dL). METHODOLOGY: Thirty-nine participants (n = 39) with ADHD, aged 7-9 years (79% male) provided saliva at 3 points during 90-minute sessions; (i) upon arrival, (ii) +20 minutes, and (iii) 15 minutes prior to departure, on 3 occasions across an 8-week intervention (weeks 1, 4, and 8). Cortisol slopes calculated within each session were compared across the intervention weeks to determine within subject and between group effect sizes. Spearmans correlations between baseline individual neurodevelopmental symptoms and in-session acute cortisol responses were also evaluated. RESULTS: No significant between group differences were observed in cortisol responsiveness at week-1. By week-4, in-session changes in cortisol were evident, with significantly greater decreases in the AAI group (Cohens d = -.40). This pattern was also observed at week-8, with an even stronger effect-size (d = -0.60). Concurrent symptoms of autism were associated with the in-session acute cortisol response. Specifically, higher parent-reported symptom scores were associated with steeper decreases in cortisol across the session at week 1 (r = -0.42, p <.01) and week-8 (r = -0.34 p = .05). At week-8 this association was stronger in the AAI group (r = -0.53) versus TAU (r = -0.25), with Cohens q = 0.413). DISCUSSION: AAI may influence acute HPA reactivity and regulation for children with ADHD. Concurrent symptoms of ADHD and autism may be related to individual differences in the nature of the effect. Implications of these findings for AAI as an alternative, or complementary intervention for ADHD are discussed. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, identifier NCT05102344.